Hardy R R, Carmack C E, Shinton S A, Riblet R J, Hayakawa K
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111.
J Immunol. 1989 May 15;142(10):3643-51.
By establishing hybridomas from two distinct surface IgM+ splenic B cell populations, Ly-1 B cells and "conventional" (Ly-1-) B cells, we found that the Ly-1 B population includes a 30 to 70 times higher frequency (1 to 2%) of cells with specificity for bromelain treated autologous red blood cells (anti-BrMRBC) when compared with conventional B cells (0.03%). We cloned and sequenced the V genes encoding anti-BrMRBC antibody from two hybridomas made with Ly-1 B cells sorted from the spleen of SM/J mice. The VH sequence (for both) is identical with the previously reported sequence associated with this specificity and belongs to a new VH gene family. This gene family, defined here as VH11, has only two members and is the predominant VH rearranged in a collection of Ly-1 B derived anti-BrMRBC hybridomas, always in association with a single VL gene (a member of the V kappa 9 family). Furthermore, analysis of hybridomas made with Ly-1 B cells sorted from the peritoneum reveals a yet higher increased frequency of VH11-encoded anti-BrMRBC specificity (30%). This variation in frequency of anti-BrMRBC in the Ly-1 population depending on location, together with the repeated association of VH11 with a particular V kappa gene suggest that antigen driven selection is (at least in part) responsible for the biased V gene expression seen in this population. Furthermore, a mechanism that might contribute to biased expression, preferential rearrangement due to close proximity to J (as seen in pre-B lines), is excluded by localization of VH11 5' to several of the more J-proximal families (Q52, 7183).
通过从两个不同的表面IgM⁺脾B细胞群体,即Ly-1 B细胞和“传统”(Ly-1⁻)B细胞建立杂交瘤,我们发现与传统B细胞(0.03%)相比,Ly-1 B细胞群体中对菠萝蛋白酶处理的自体红细胞具有特异性(抗BrMRBC)的细胞频率高30至70倍(1%至2%)。我们从用SM/J小鼠脾脏中分选的Ly-1 B细胞制备的两个杂交瘤中克隆并测序了编码抗BrMRBC抗体的V基因。VH序列(两者)与先前报道的与此特异性相关的序列相同,属于一个新的VH基因家族。这个基因家族在此定义为VH11,只有两个成员,是Ly-1 B衍生的抗BrMRBC杂交瘤集合中重排的主要VH基因,总是与单个VL基因(Vκ9家族的一个成员)相关联。此外,对用从腹膜中分选的Ly-1 B细胞制备的杂交瘤的分析显示,VH11编码的抗BrMRBC特异性频率更高(30%)。Ly-1群体中抗BrMRBC频率因位置而异,以及VH11与特定Vκ基因的反复关联表明,抗原驱动的选择(至少部分)是该群体中V基因表达偏向的原因。此外,一种可能导致表达偏向的机制,即由于靠近J而优先重排(如在前B细胞系中所见),被VH11 5'定位到几个更靠近J的家族(Q52、7183)所排除。